Herein, the objectives of this study are to (1) investigate the prognostic value of PUS1 gene expressions in HCC using datasets from The Cancer Genome Atlas (TCGA), the Clinical Proteomic Tumor Analysis Consortium (CPTAC), the UALCAN website, THE HUMAN PROTEIN ATLAS, the TCGA-HCC database, and HCC patients’ tumor tissues; (2) investigate the potential mechanism of PUS1 affecting occurrence and development of HCC; and (3) verify the mechanism based on experiments on HCC cell lines. This evidence concerns the gene PUS1 and neoplasm.